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Updated: Aug 15, 2026
Murine Dermal Lymphatic Endothelial Cell Isolation
Published on: July 21, 2023
A method of isolation and culture of microvascular endothelial cells from mouse skin
Sung Tae Cha1, Dodanim Talavera, Erhan Demir
1Department of Surgery, Division of Plastic and Reconstructive Surgery, Engineered Wound Repair Laboratory at Davis Building, Cedars Sinai Medical Center, 8700 Beverly Blvd., Los Angeles, CA 90048, USA.
Objectives:
The study of isolated microvascular endothelial cells from mice has long been impeded due to the many difficulties encountered in isolating and culturing these cells. We focused on developing a method to isolate microvascular endothelial cells from the skin fragments of newborn mice. We also aimed at establishing optimal culture conditions to sustain the growth of these cells.
Methods And Results:
Isolation of murine dermal microvascular endothelial cells (mDMEC) from P3 newborn mice was based first on enzymatic separation of the skin epidermal layer from the dermis using dispase and then on disaggregating dermal cellular elements using collagenase. The cells obtained from the dermis were subjected to a continuous density gradient centrifugation. Cells situated between densities 1.033 and 1.047 were then cultured on collagen IV-coated culture flasks using optimized growth culture conditions. Cells were characterized by endothelial appearance and by the presence and genetic expression of endothelial markers like CD31, NOS3, VEGFR-2 and Tie-2. Uptake of acetylated low-density lipoprotein (Ac-LDL) was used as a functional assay.
Conclusions:
The methodology described herein for isolation and culture of murine microvascular endothelium offers a distinctive advantage for those using mouse models to study endothelial cell biology.
Insights
Researchers developed a new method to isolate and culture murine dermal microvascular endothelial cells (mDMEC) from newborn mice. This technique simplifies studying endothelial cell biology in mouse models.
Area of Science:
- Cell Biology
- Microcirculation Research
- Endothelial Cell Biology
Background:
- Isolating and culturing microvascular endothelial cells from mice presents significant challenges.
- Existing methods are often difficult and time-consuming, limiting research progress.
Purpose of the Study:
- To develop an efficient method for isolating microvascular endothelial cells from newborn mouse skin.
- To establish optimal culture conditions for sustaining the growth of these cells.
Main Methods:
- Skin epidermal layer separation using dispase, followed by dermal cell disaggregation with collagenase.
- Density gradient centrifugation to isolate cells, followed by culture on collagen IV-coated flasks.
- Characterization using endothelial markers (CD31, NOS3, VEGFR-2, Tie-2) and acetylated low-density lipoprotein uptake assay.
Main Results:
- Successfully isolated murine dermal microvascular endothelial cells (mDMEC) from P3 newborn mice.
- Established optimal culture conditions supporting cell growth and viability.
- Confirmed cell identity through morphological and molecular markers, including CD31, NOS3, VEGFR-2, and Tie-2 expression.
Conclusions:
- The described methodology provides a reliable approach for isolating and culturing murine microvascular endothelial cells.
- This technique offers a significant advantage for researchers utilizing mouse models in endothelial cell biology studies.
